目的综述分化抑制因子2(inhibitor of differentiation2,Id2)在骨骼肌再生中的作用研究进展。方法广泛查阅近年来Id2生物学特性及其在骨骼肌再生中的作用相关文献,并综合分析。结果Id2通过结合E蛋白形成异二聚体,阻止生肌调节因子(myoge...目的综述分化抑制因子2(inhibitor of differentiation2,Id2)在骨骼肌再生中的作用研究进展。方法广泛查阅近年来Id2生物学特性及其在骨骼肌再生中的作用相关文献,并综合分析。结果Id2通过结合E蛋白形成异二聚体,阻止生肌调节因子(myogenic regulatory factors,MRFs)与E蛋白结合而抑制MRFs的转录活性,抑制骨骼肌细胞分化。结论Id2在骨骼肌再生中起着重要作用。展开更多
AIM: To investigate the effect of herbal compound 861 (Cpd861) on the transforming growth factor-β1 (TGFβ1)/ activin receptor-like kinase 1 (ALK1, type Ⅰ receptor) signaling-pathway-related gene expression in the L...AIM: To investigate the effect of herbal compound 861 (Cpd861) on the transforming growth factor-β1 (TGFβ1)/ activin receptor-like kinase 1 (ALK1, type Ⅰ receptor) signaling-pathway-related gene expression in the LX-2 cell line, and the inhibitory mechanism of Cpd861 on the activation of LX-2 cells. METHODS: LX-2 cells were treated with TGFβ1 (5 ng/mL) Cpd861 (0.1 mg/mL), TGFβ1 (5 ng/mL) plus Cpd861 (5 ng/mL) for 24 h to investigate the effect of Cpd861 on the TGFβ1/ALK1 pathway. Real-time PCR was performed to examine the expression of α-SMA (α-smooth muscle actin), ALK1, Id1 (inhibitor of differentiation 1). Western blotting was carried out to measure the levels of α-SMA and phosphorylated Smad1, and immunocytochemical analysis for the expression of α-SMA. RESULTS: In LX-2 cells, TGFβ1/ALK1-pathway-related gene expression could be stimulated by TGFβ1, which led to excessive activation of the cells. Cpd861 decreased the activation of LX-2 cells by reducing the expression of α-SMA mRNA and protein expression. This effect was related to inhibition of the above TGFβ1/ALK1-pathway- related expression of genes such as Id1 and ALK1, and phosphorylation of Smad1 in LX-2 cells, even with TGFβ1 co-treatment for 24 h. CONCLUSION: Cpd861 can restrain the activation of LX-2 cells by inhibiting the TGFβ1/ALK1/Smad1 pathway.展开更多
Progression,relapse,and therapy resistance are the most challenging features of cancer therapy that have been postulated to be driven by Cancer Stem Cell(CSC)population.This enigmatic subpopulation of cancer cells has...Progression,relapse,and therapy resistance are the most challenging features of cancer therapy that have been postulated to be driven by Cancer Stem Cell(CSC)population.This enigmatic subpopulation of cancer cells has therefore emerged as promising therapeutic candidate.We earlier reported enrichment of CSC-like side population(SP)with increasing resistance towards Cisplatin and Paclitaxel either alone or in combination in epithelial ovarian cancer(EOC)cells.This SP population is a small proportion of the total population of cancer cells characterised with high expression of drug transporters,a unique feature of stem cells and thereby can be isolated through their efflux properties of DNA binding dyes.While the bulk non-SP(NSP)population of the cancer cells lack overexpression of the drug transporters and thus can be identified as the dye containing population.In this study,we show that increased expression of Runt related transcription factor 1(RUNX1)maintains undifferentiated state of CSC-like SP cells through upregulation of inhibitors of DNA binding/differentiation genes(ID1 and ID3)in late cisplatinpaclitaxel resistant cells.Higher RUNX1 expression was found to correlate with decreased median overall survival and disease-free survival in The Cancer Genome Atlas(TCGA)data set of high grade serous ovarian cancer(HGSOC)patients.The protein-protein interaction network analysis of 397 upregulated genes in RUNX1-high samples of TCGA data show significant enrichment of pathways known to negatively regulate CSC differentiation.Intriguingly RUNX1 inhibition not only induces CSC differentiation but also downregulates anti-apoptotic protein BCL2 in both SP and NSP cells and potentiates cytotoxic effects of Cisplatin-Paclitaxel in chemoresistant EOC cells.Inhibition of BCL2 through Venetoclax treatment,a small molecule BH3 mimic,sensitized these cells to platinum taxol treatment.Altogether,our data reveal new regulatory roles by RUNX1 to modulate CSC differentiation via ID1 and ID3 and to promote chemoresistance through BCL2 upregulation.展开更多
目的构建转录因子E2-2基因腺病毒载体,观测内皮祖细胞(endothelial progenitor cells,EPCs)过表达E2-2基因对DNA结合抑制因子-1(inhibitor of DNA binding/differentiation,ID1)表达的影响。方法分离、培养并鉴定小鼠骨髓EPCs。RT-PCR...目的构建转录因子E2-2基因腺病毒载体,观测内皮祖细胞(endothelial progenitor cells,EPCs)过表达E2-2基因对DNA结合抑制因子-1(inhibitor of DNA binding/differentiation,ID1)表达的影响。方法分离、培养并鉴定小鼠骨髓EPCs。RT-PCR法扩增E2-2基因CDs全长DNA,克隆入载体pTG19-T后,亚克隆入腺病毒穿梭载体pAdTrack-CMV中,构建pAdTrack/E2-2重组载体,与pAdEasy-1骨架质粒同源重组形成重组病毒pAd/E2-2,经293细胞包装,获具高效感染力的重组pAd/E2-2病毒。将该病毒感染EPCs,倒置显微镜观测经感染的EPCs的GFP表达情况。CCK-8(cell count kit-8)法检测病毒pAd/E2-2对EPCs生长、增殖的影响。RT-PCR、Western blot分别检测经感染的EPCs中E2-2与ID1基因及其编码蛋白的表达情况,并予以定量分析。结果分离、培养并鉴定到小鼠骨髓EPCs。克隆到2013 bp的E2-2基因,并获得高效感染力的重组pAd/E2-2病毒。CCK-8法检测表明,与对照比较,过表达E2-2的EPCs的生长、增殖速度减慢,48h开始变得尤为明显(P<0.01);RT-PCR、Western blot及定量分析结果显示,E2-2能下调ID1的表达,与对照比较,差异具统计学意义(P<0.01)。结论分离、培养并鉴定小鼠骨髓EPCs,克隆出E2-2基因,证实E2-2能明显抑制EPCs的生长、增殖,并能下调ID1基因的表达。展开更多
目的研究分化抑制因子-2(inhibitor of differentiation-2,Id-2)和上皮性钙黏附素(epithelical cadherin,E-cadherin)在结肠癌组织中的表达及其相关性。方法采用HE染色观察癌旁正常组织、癌组织的病理改变情况;免疫组织化学方法检测Id-2...目的研究分化抑制因子-2(inhibitor of differentiation-2,Id-2)和上皮性钙黏附素(epithelical cadherin,E-cadherin)在结肠癌组织中的表达及其相关性。方法采用HE染色观察癌旁正常组织、癌组织的病理改变情况;免疫组织化学方法检测Id-2和Ecadherin在癌旁正常组织、癌组织中的表达水平,Spearman相关检验检测Id-2与E-cadherin表达的相关性。结果 HE染色结果:结肠癌组织:癌细胞分化差,多形性,大小不一。可呈假复层,核大,胞浆少,容易找到核分裂。免疫组织化学检测结果:Id-2在癌旁正常黏膜组织和结肠癌组织的阳性表达率分别为57.5%和82.5%;E-cadherin的阳性表达率分别为72.5%和42.5%。两组Id-2和E-cadherin的阳性表达率比较,差异有统计学意义(P<0.05)。Spearman相关分析:Id-2与E-cadherin的表达水平呈负相关性(P<0.05)。结论结肠癌组织中Id-2高表达可能通过E-cadherin促使肿瘤组织细胞发生侵袭、转移。展开更多
文摘目的综述分化抑制因子2(inhibitor of differentiation2,Id2)在骨骼肌再生中的作用研究进展。方法广泛查阅近年来Id2生物学特性及其在骨骼肌再生中的作用相关文献,并综合分析。结果Id2通过结合E蛋白形成异二聚体,阻止生肌调节因子(myogenic regulatory factors,MRFs)与E蛋白结合而抑制MRFs的转录活性,抑制骨骼肌细胞分化。结论Id2在骨骼肌再生中起着重要作用。
文摘AIM: To investigate the effect of herbal compound 861 (Cpd861) on the transforming growth factor-β1 (TGFβ1)/ activin receptor-like kinase 1 (ALK1, type Ⅰ receptor) signaling-pathway-related gene expression in the LX-2 cell line, and the inhibitory mechanism of Cpd861 on the activation of LX-2 cells. METHODS: LX-2 cells were treated with TGFβ1 (5 ng/mL) Cpd861 (0.1 mg/mL), TGFβ1 (5 ng/mL) plus Cpd861 (5 ng/mL) for 24 h to investigate the effect of Cpd861 on the TGFβ1/ALK1 pathway. Real-time PCR was performed to examine the expression of α-SMA (α-smooth muscle actin), ALK1, Id1 (inhibitor of differentiation 1). Western blotting was carried out to measure the levels of α-SMA and phosphorylated Smad1, and immunocytochemical analysis for the expression of α-SMA. RESULTS: In LX-2 cells, TGFβ1/ALK1-pathway-related gene expression could be stimulated by TGFβ1, which led to excessive activation of the cells. Cpd861 decreased the activation of LX-2 cells by reducing the expression of α-SMA mRNA and protein expression. This effect was related to inhibition of the above TGFβ1/ALK1-pathway- related expression of genes such as Id1 and ALK1, and phosphorylation of Smad1 in LX-2 cells, even with TGFβ1 co-treatment for 24 h. CONCLUSION: Cpd861 can restrain the activation of LX-2 cells by inhibiting the TGFβ1/ALK1/Smad1 pathway.
文摘Progression,relapse,and therapy resistance are the most challenging features of cancer therapy that have been postulated to be driven by Cancer Stem Cell(CSC)population.This enigmatic subpopulation of cancer cells has therefore emerged as promising therapeutic candidate.We earlier reported enrichment of CSC-like side population(SP)with increasing resistance towards Cisplatin and Paclitaxel either alone or in combination in epithelial ovarian cancer(EOC)cells.This SP population is a small proportion of the total population of cancer cells characterised with high expression of drug transporters,a unique feature of stem cells and thereby can be isolated through their efflux properties of DNA binding dyes.While the bulk non-SP(NSP)population of the cancer cells lack overexpression of the drug transporters and thus can be identified as the dye containing population.In this study,we show that increased expression of Runt related transcription factor 1(RUNX1)maintains undifferentiated state of CSC-like SP cells through upregulation of inhibitors of DNA binding/differentiation genes(ID1 and ID3)in late cisplatinpaclitaxel resistant cells.Higher RUNX1 expression was found to correlate with decreased median overall survival and disease-free survival in The Cancer Genome Atlas(TCGA)data set of high grade serous ovarian cancer(HGSOC)patients.The protein-protein interaction network analysis of 397 upregulated genes in RUNX1-high samples of TCGA data show significant enrichment of pathways known to negatively regulate CSC differentiation.Intriguingly RUNX1 inhibition not only induces CSC differentiation but also downregulates anti-apoptotic protein BCL2 in both SP and NSP cells and potentiates cytotoxic effects of Cisplatin-Paclitaxel in chemoresistant EOC cells.Inhibition of BCL2 through Venetoclax treatment,a small molecule BH3 mimic,sensitized these cells to platinum taxol treatment.Altogether,our data reveal new regulatory roles by RUNX1 to modulate CSC differentiation via ID1 and ID3 and to promote chemoresistance through BCL2 upregulation.
文摘目的研究分化抑制因子-2(inhibitor of differentiation-2,Id-2)和上皮性钙黏附素(epithelical cadherin,E-cadherin)在结肠癌组织中的表达及其相关性。方法采用HE染色观察癌旁正常组织、癌组织的病理改变情况;免疫组织化学方法检测Id-2和Ecadherin在癌旁正常组织、癌组织中的表达水平,Spearman相关检验检测Id-2与E-cadherin表达的相关性。结果 HE染色结果:结肠癌组织:癌细胞分化差,多形性,大小不一。可呈假复层,核大,胞浆少,容易找到核分裂。免疫组织化学检测结果:Id-2在癌旁正常黏膜组织和结肠癌组织的阳性表达率分别为57.5%和82.5%;E-cadherin的阳性表达率分别为72.5%和42.5%。两组Id-2和E-cadherin的阳性表达率比较,差异有统计学意义(P<0.05)。Spearman相关分析:Id-2与E-cadherin的表达水平呈负相关性(P<0.05)。结论结肠癌组织中Id-2高表达可能通过E-cadherin促使肿瘤组织细胞发生侵袭、转移。
